Chlamydia MODULE

Microbiology

33

CHLAMYDIA Notes

33.1 INTRODUCTION
Chlamydiae are obligate, aerobic, intracellular parasites of eukaryotic cells.
They are small Gram-negative coccoid or rod shaped, non-motile bacteria.
Chlamydiae exhibit characteristics intermediate between bacteria and viruses.
They are widespread in the natural world, being parasites of people, animals
and birds with tropism for squamous epithelial cells and macrophages of the
respiratory and gastrointestinal tract.
They are recognized as bacteria as
z They have both DNA and RNA.
z They have cell wall (that resembles that of GNB) and ribosomes
z Replicate by binary fission
z Susceptible to antibiotics

OBJECTIVES
After reading this lesson, you will be able to:
z distinguish characteristics of Chlamydia.
z describe the diseases produced by it.
z explain the laboratory diagnosis.

33.2 CLASSIFICATION
Classification of Chlamydiae is as follows:
Order Chlamydiales

30 MICROBIOLO MICROBIOLO 30
 MODULE Chlamydia

Microbiology Family Chlamydiaceae

Genus Chlamydia,Chlamydophila
Species There are three species C. trachomatis, Chlamydophila psittaci,
Chlamydophila pneumoniaae
C. trachomatis has two biovars; TRIC and LGV.
Notes
33.3 CELL STRUCTURE
Chlamydiae have a cytoplasmic membrane and an outer membrane similar to
Gram-negative bacteria but lack a peptidoglycan cell wall. Chlamydiae cannot
synthesize their own ATP and require intracellular abode to remain viable.
Chlamydiae exist in two forms: the elementary body and the reticulate body.
Both of them play a pivotal part in the life cycle of chlamydia. Although Gram
negative, Chlamydiae stain better with Castaneda, Machiavello or Gimenez
stains.

33.4 ELEMENTARY BODY (EB)

The elementary body is the dispersal form, which is analogous to a spore. This
dispersal form is about 0.3 µm or 200-300 nm in diameter. It is the
extracellular infective form. It induces its own endocytosis upon exposure to
target cells.

33.5 RETICULATE BODY (RB)

Reticulate body is the intracellular, multiplicative form. It represents the non-
infectious growing form.

33.6 LIFE CYCLE

The life cycle of Chlamydia trachomatis consists of two stages: elementary
body and reticulate body. Upon endocytosis into the host cell EB prevents
phagolysosomal fusion enabling intracellular survival of the bacteria. Once
inside the endosome, the elementary body transforms into the larger reticulate
body (500 – 1000 nm) as a result of the glycogen that is produced. The
reticulate body is the reproductive form. It divides through binary fission at
approximately
2-3 hours per generation. It contains no cell wall and is detected as an
inclusion in the cell arranged as a mantle around the nucleus. The inclusion
bodies are basophilic. They can also be stained by Lugol’s iodine because of
the presence of glycogen matrix. After division, the reticulate body
transforms back to the elementary form and is released by the cell by
exocytosis. One phagolysosome usually produces 100-1000 elementary

31 MICROBIOLO MICROBIOLO 31
bodies. The entire
process takes 24 – 48
hours. The EB may
infect new cells and
the cycle continues.

31 MICROBIOLO MICROBIOLO 31
 Chlamydia MODULE
Microbiology
33.7 ANTIGENIC STRUCTURE
Chlamydia antigens consist of 3 groups: genus-specific antigen, species-
specific protein antigen, serotype-specific. The sero type-specific antigens are
located on MOMP and on the basis of this chlamydiae have been divided into
many serovars or serotypes.

33.8 CULTURE Notes

Chlamydiae can be isolated by the following methods:
(a) Animal inoculation: Mice can be inoculated through intranasal,
intraperitoneal or intracerebral route. Mice die within 10 days. Smears
made from lung, spleen, brain or peritoneal exudate demonstrate
elementary bodies.
(b) Egg inoculation: Organisms can be isolated by egg yolk inoculation of
the specimen. Impression smears can be stained by Giemsa or Gimenez.
(c) Tissue culture: McCoy cells treated with cycloheximide are the most
commonly used cell lines. Irradiated or metabolically inhibited cell lines
can also be used for isolation of chlamydia. Inclusion bodies can be
visualized by staining the cell lines.

33.9 DISEASES PRODUCED BY CHLAMYDIA

(a) Ocular infections: C. trachomatis serotype A,B,Ba,C- is the leading
cause of preventable blindness (caused by a chlamydia infection called
trachoma) in the world. Other diseases produced are inclusion
conjunctivitis (serotype D to K) and ophthalmia neonatorum.
(b) Genital infections: C. trachomatis is also the leading cause of sexually
transmitted disease worldwide. It is associated with non- gonococcal
urethritis and lymphogranuloma venereum (serotype L1, L2, L3). C.
trachomatis is one of the major causes of pelvic inflammatory disease
(PID) and infertility in women.
(c) Respiratory infections: C. pneumoniae causes pneumonia. C. psittaci
causes psittacosis.

33.10 LABORATORY DIAGNOSIS

Specimen collection: Specimen should be collected by scraping the mucosa.
Discharge should not be collected. Depending on the site of infection, ocular,
urethral, cervical, sputum, respiratory secretions can be collected. In suspected
Psittacosis, blood and sputum are collected for microscopy and culture and
serum for serology.

31 MICROBIOLO MICROBIOLO 31
 MODULE Chlamydia

Microbiology Direct detection of antigen: Antigen detection is a rapid method of diagnosing

chlamydial infection.
1. Light Microscopy: Inclusion bodies of C. trachomatis can be detected by
staining with Lugol’s iodine. Iodine can be used because inclusion bodies
contain a glycogen matrix. Giemsa, Castaneda, Machiavello and Giminez
methods are better and can be used to stain ocular, cervical or urethral
specimen.
Notes 2. Immunofluoresence: Direct fluorescent antibody test detects major outer
membrane proteins. It is now considered by many the test of choice for
diagnosis.

ELISA: Antigen and antibodies can be detected by ELISA. Antigen detection

is more specific than antibody detection.
Isolation: Mice, fertilized hen’s egg and tissue cultures can be used for
isolation of chlamydia. The clinical specimen can be inoculated into the yolk
sac of 6 to
8 day old eggs. Irradiated or cycloheximide treated McCoy cell culture is the
preferred isolation method.
Molecular tools: Polymerase chain reaction, ligase chain reaction can be used
for detection of chlamydia

33.11 TREATMENT
Sulphonamides and tetracycline are the drugs of choice. Single dose
azithromycin is the drug of choice for non-gonoccocal urethritis.

INTEXT QUESTIONS 33.1

1. Chlamydiae are ............... parasites
2. Chlamydiae are gram ............... cocci
3. ............... body enables intracellular survival of the bacteria
4. ............... body is the reproductive form
5. C.trahomatis causes ............... & ...............
31 MICROBIOLO MICROBIOLO 31
 Chlamydia MODULE
Microbiology

WHAT YOU HAVE LEARNT

z Chlamydiae are obligate, aerobic, intracellular parasites of eukaryotic
cells. z They are small Gram-negative coccoid or rod shaped, non-motile
bacteria. z Chlamydiae has elementary body which is similar to a spore.
z Life cycle of Chlamydia trachomatis consists of two stages namely Notes
elementary and reticulate body.
z Chlamydiae can be isolated by animal inoculation, egg inoculation &
tissue culture
z Chlamydia trachomatis causes preventable blindness, conjunctivitis &
ophthalmia neonatorum, non- gonococcal urethritis and lymphogranuloma
venereum, inflammatory disease, infertility in women.
z Inclusion bodies of C. trachomatis can be detected by staining with Lugol's
iodine
z Giemsa, Castaneda, Machiavello methods are used to stain ocular, cervical
or urethral specimen.
z Direct fluorescent antibody is test of choice for diagnosis.

TERMINAL EXERCISES
1. Describe the life cycle of chlamydia.
2. Name the various staining techniques used for chlamydia.
3. Enumerate the diseases caused by Chlamydia trachomatis.
4. How can we cultivate chlamydia?
5. What are the rapid diagnostic methods for detecting chlamydia?

ANSWERS TO INTEXT QUESTIONS

33.1
1. Intracellular
2. Negative
3. Elementary
4. Reticulate
5. Blindness & lymphogranuloma venerum

31 MICROBIOLO MICROBIOLO 31